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IMPROVE SILENCING SUCCESS: siRNAs THAT WORK BETTER!


Effectiveness of Silencer Validated siRNAs
As more is learned about siRNAs, it is becoming increasingly clear that using highly potent siRNAs that efficiently silence their target gene at low siRNA concentrations is advantageous. Transfection of low siRNA concentrations reduces the potential for off target effects or induction of the antiviral response. Figure 2 shows data from six Silencer Validated siRNAs tested at five different concentrations. In all cases, silencing efficiency at 10 nM was comparable to that achieved at 100 nM siRNA, indicating that these siRNAs are effective at low concentrations. Three of these siRNAs were tested further by transfecting HepG2, SK-N-AS, and MCF7 cells at five different concentrations (Figure 3). Even in these more difficult-to-transfect cells, the Silencer Validated siRNAs were effective at low concentrations.

Figure 2. Silencer Validated siRNAs Are Effective at Low Concentrations. The indicated siRNAs were transfected into HeLa cells using siPORT Lipid at the final siRNA concentrations shown. Forty-eight hours after transfection, RNA was recovered using RNAqueous MAG-96 RNA Isolation Kit and reverse transcribed using the RETROscript Kit. Target cDNA levels were measured by real-time PCR and compared to cells transfected with an equal concentration of Silencer Negative Control #1. cDNA was normalized using real-time data for 18S rRNA. The bar graphs represent an average of three data points.

Figure 3. Silencer Validated siRNA Activity in Various Cell Types. HeLa, HepG2, MCF7, and SK-N-AS cells in 24 well plates were transfected with the indicated siRNAs at
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